A systematic characterization of Cwc21, the yeast ortholog of the human spliceosomal protein SRm300

May Khanna, Harm Van Bakel, Xinyi Tang, John A. Calarco, Tomas Babak, Grace Guo, Andrew Emili, Jack F. Greenblatt, Timothy R. Hughes, Nevan J. Krogan, Benjamin J. Blencowe

Research output: Contribution to journalArticle

26 Scopus citations

Abstract

Cwc21 (complexed with Cef1 protein 21) is a 135 amino acid yeast protein that shares homology with the N-terminal domain of human SRm300/SRRM2, a large serine/arginine-repeat protein shown previously to associate with the splicing coactivator and 39-end processing stimulatory factor, SRm160. Proteomic analysis of spliceosomal complexes has suggested a role for Cwc21 and SRm300 at the core of the spliceosome. However, specific functions for these proteins have remained elusive. In this report, we employ quantitative genetic interaction mapping, mass spectrometry of tandem affinity-purified complexes, and microarray profiling to investigate genetic, physical, and functional interactions involving Cwc21. Combined data from these assays support multiple roles for Cwc21 in the formation and function of splicing complexes. Consistent with a role for Cwc21 at the core of the spliceosome, we observe strong genetic, physical, and functional interactions with Isy1, a protein previously implicated in the first catalytic step of splicing and splicing fidelity. Together, the results suggest multiple functions for Cwc21/SRm300 in the splicing process, including an important role in the activation of splicing in association with Isy1. Published by Cold Spring Harbor Laboratory Press.

Original languageEnglish (US)
Pages (from-to)2174-2185
Number of pages12
JournalRNA
Volume15
Issue number12
DOIs
StatePublished - Dec 1 2009
Externally publishedYes

Keywords

  • Cwc21
  • SRm300
  • SnRNP
  • Spliceosome
  • Splicing

ASJC Scopus subject areas

  • Molecular Biology

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    Khanna, M., Van Bakel, H., Tang, X., Calarco, J. A., Babak, T., Guo, G., Emili, A., Greenblatt, J. F., Hughes, T. R., Krogan, N. J., & Blencowe, B. J. (2009). A systematic characterization of Cwc21, the yeast ortholog of the human spliceosomal protein SRm300. RNA, 15(12), 2174-2185. https://doi.org/10.1261/rna.1790509